Meeting report: Spontaneous lesions and diseases in wild, captive-bred, and zoo-housed nonhuman primates and in nonhuman primate species used in drug safety studies.

The combination of loss of habitat, human population encroachment, and increased demand of select nonhuman primates for biomedical research has significantly affected populations. There remains a need for knowledge and expertise in understanding background findings as related to the age, source, strain, and disease status of nonhuman primates. In particular, for safety/biomedical studies, a broader understanding and documentation of lesions would help clarify background from drug-related findings. A workshop and a minisymposium on spontaneous lesions and diseases in nonhuman primates were sponsored by the concurrent Annual Meetings of the American College of Veterinary Pathologists and the American Society for Veterinary Clinical Pathology held December 3-4, 2011, in Nashville, Tennessee. The first session had presentations from Drs Lowenstine and Montali, pathologists with extensive experience in wild and zoo populations of nonhuman primates, which was followed by presentations of 20 unique case reports of rare or newly observed spontaneous lesions in nonhuman primates (see online files for access to digital whole-slide images corresponding to each case report at http://www.scanscope.com/ACVP%20Slide%20Seminars/2011/Primate%20Pathology/view.apml). The minisymposium was composed of 5 nonhuman-primate researchers (Drs Bradley, Cline, Sasseville, Miller, Hutto) who concentrated on background and spontaneous lesions in nonhuman primates used in drug safety studies. Cynomolgus and rhesus macaques were emphasized, with some material presented on common marmosets. Congenital, acquired, inflammatory, and neoplastic changes were highlighed with a focus on clinical, macroscopic, and histopathologic findings that could confound the interpretation of drug safety studies.

Novel endotheliotropic herpesviruses fatal for Asian and African elephants.

A highly fatal hemorrhagic disease has been identified in 10 young Asian and African elephants at North American zoos. In the affected animals there was ultrastructural evidence for herpesvirus-like particles in endothelial cells of the heart, liver, and tongue. Consensus primer polymerase chain reaction combined with sequencing yielded molecular evidence that confirmed the presence of two novel but related herpesviruses associated with the disease, one in Asian elephants and another in African elephants. Otherwise healthy African elephants with external herpetic lesions yielded herpesvirus sequences identical to that found in Asian elephants with endothelial disease. This finding suggests that the Asian elephant deaths were caused by cross-species infection with a herpesvirus that is naturally latent in, but normally not lethal to, African elephants. A reciprocal relationship may exist for the African elephant disease.

Extrahepatic biliary carcinomas in Asiatic bears.

Extrahepatic biliary carcinomas occurred in 2 Asiatic bears at the National Zoological Park in Washington, D.C. The tumors were associated with ascites in both cases and jaundice in 1 and metastasized widely. Neither bear had gallstones. Microscopically, the tumors were infiltrative, scirrhous adenocarcinomas that produced varying amounts of mucin. A genetic predisposition and change from the natural diet were factors proposed to contribute to the development of this neoplasm.

Familial goiter in bongo antelope (Tragelaphus eurycerus).

Congenital defects in thyroglobulin (Tg) synthesis in animals have proven to be useful models for the study of Tg synthesis and regulation. Defects in Tg synthesis have been well described in Afrikander cattle, Australia Merino sheep, and goats in The Netherlands. This report describes a study of goiter in a nondomesticated bovine species, bongo antelope (Tragelaphus eurycerus), an African bovid. Three animals housed at the National Zoological Park, Washington, D.C. were studied; two had visible goiters, and a third bongo had microscopic evidence of goiter. Tg extracted from thyroid glands or thyroid colloid from these animals had a high mol wt component that was greater than 220K daltons and differed in apparent mol wt from 19S Tg from domestic cattle. Thyroid extracts also had thyroid albumin; albumin was more than half the total protein in colloid extract. The animals with goiter were euthyroid according to their circulating levels of thyroid hormones.

Natural history of autologous blood clot embolization in swine.

Previous studies of the natural history of embolized clots in dogs have demonstrated rapid lysis, presumably because the canine fibrinolytic system is very active. The fibrinolytic activity in swine, however, is similar to humans, and for this reason the pig was chosen for our study. The gluteal branches of the external iliac artery in nine domestic swine were embolized with either unmodified or modified (heat-formed, Amicar) autologous clot. In addition, three pigs were embolized with unmodified autologous clot to branches of the gastrosplenic artery. The lysis of clot emboli in both groups was followed by serial angiography at 48 hours and 14 days. Clot lysis as assessed by euglobulin lysis and plasmin generation was not activated by the experimental technique. Necropsy was performed on the animals in the second group. Partial or total obstruction of all arteries was present 48 hours after embolization and only 50% of arteries were recanalized at 14 days. At necropsy, organized partially occluding clot was demonstrated in the splenic artery of all 3 embolized swine. It is concluded that: 1)swine provide an excellent animal model for studying the natural history of arterial embolization; 2)Amicar or heat-formed clot shows no advantage over simple autologous clot in retarding intra-arterial clot lysis, and 3)simple autologous clot is an effective material for temporary intra-arterial occlusion.

A variant of the endotheliotropic herpesvirus in Asian elephants (Elephas maximus) in European zoos.

Newly discovered, lethal elephant endotheliotropic herpesviruses (EEHV) have been identified in both Asian (Elephas maximus) and African (Loxodonta africana) elephants. Carried by otherwise healthy African elephants they can be fatal mainly for young Asian elephants. Since zoos often harbour both elephant species, we conducted a survey on the presence of EEHV in Asian elephants from 12 European zoos, 3 circuses and 1 Israeli zoo. Here, we demonstrate that all EEHV that have affected Asian elephants so far belong to the EEHV1 group. We also describe the detection and the partial sequencing of an endotheliotropic herpesvirus variant (named EEHV1b) in Asian elephants, being either an EEHV endogenous to Asian elephants or indicating different sources (African elephants) of infection.

Mycobacterium marinum dermatitis and panniculitis with chronic pleuritis in a captive white whale (Delphinapterus leucas) with aortic rupture.

A 16-year-old female white whale, Delphinapterus leucas, died after nearly 18 months of chronic lymphopenia and pyogranulomatous dermatitis. Necropsy revealed rupture of the aorta with hemorrhage into the cranial mediastinum and between fascial planes of the ventral neck musculature. Multiple foci of ulcerative dermatitis and panniculitis were present across the thorax and abdomen and surrounded the genital folds. In addition, there was a chronic proliferative pleuritis with over 20 liters of histiocytic exudate in the thoracic cavity. Acid-fast bacteria consistent with Mycobacterium sp. were identified in sections of skin lesions and in cytospins of pleural exudate. Cultures of pleura and 1 skin lesion collected at necropsy yielded sparse growth of an acid-fast bacillus with colony characteristics and morphology consistent with Mycobacterium marinum. Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) analysis confirmed the presence of M. marinum DNA in samples of skin. This is the first documented occurrence of mycobacteriosis in a white whale and is a unique presentation of mycobacterial dermatitis and panniculitis with chronic pleuritis in a cetacean. The improved PCR-RFLP protocol utilized in this case unifies techniques from several protocols to differentiate between species of Nocardia and rapidly growing mycobacteria clinically relevant to aquatic animals.

Diverticulitis with rupture and fatal peritonitis in a Sumatran orangutan (Pongo pygmaeus).

A 30-year-old male Sumatran orangutan (Pongo pygmaeus) presented with signs of depression, lethargy, anorexia, and diarrhea that progressed to acute colic. Exploratory laparotomy revealed fibrinopurulent peritonitis and 50 cm of devitalized small intestine. The surgically resected small intestine contained several mucosal diverticula along the mesenteric attachment; one had ruptured, resulting in peritonitis. Fifteen days after surgery, the orangutan's abdominal incision dehisced. Repeated laparotomy revealed dehiscence of the distal intestinal anastomosis site, as well as extensive adhesions and purulent exudate. The defect was repaired, and the abdomen was extensively irrigated and closed, but the animal died within 24 hours. To our knowledge, this is the first report of diverticulitis in a great ape. Diverticulosis should be considered in the differential diagnosis for great apes that present with signs of depression, lethargy, anorexia, and/or diarrhea.

Surgical correction of familial diaphragmatic hernia of Morgagni in the Golden Lion Tamarin.

The Golden Lion Tamarin (Leontopithecus rosalia) is an endangered species of primate indigenous to the coastal rain forest of Brazil. Since 1971 a propagation and behavioral research program has been carried out on a colony of these monkeys at the Zoological Park. Several related animals have died and at necropsy have shown absence of the anterior portion of the diaphragm with a variety of abdominal viscera in the thorax. Diagnostic studies undertaken on the living members of the colony include plain chest radiographs and gastrointestinal series. Four of the animals have been found to have major diaphragmatic defects with the liver, stomach, spleen, colon, and portions of the intestine in the chest. In three of the animals so affected clinical signs of failure-to-thrive were clearly manifest, although one female born in 1968 has reproduced successfully for 8 yr with no deleterious clinical effects from the hernia. Surgical repair of the diaphragm with relocation of the abdominal viscera has been carried out in three of the animals under a combination of intravenous and intra-tracheal anesthesia. The monkeys weighed 550, 685, and 710 g, respectively. All three Tamarins have survived the operative procedure, and subsequently have shown improved clinical status. These cases have been instructive from the genetic point of view, the major anatomical defect, the type of repair necessary, and the postoperative care in small vigorous mammals. Additionally this study documents a familial factor in diaphragmatic defects in higher mammals.

Terrestrial gastropod hosts of Parelaphostrongylus tenuis at the National Zoological Park's Conservation and Research Center, Virginia.

Terrestrial gastropods at the National Zoological Park's Conservation and Research Center (CRC) near Front Royal, Virginia, were surveyed from June 1985 to May 1986, to assess their role in transmission of the meningeal worm, Parelaphostrongylus tenuis. A total of 670 gastropods representing 9 families and 18 species was collected. The slug Deroceras laeve accounted for 50.4% of the specimens collected. Parelaphostrongylus tenuis infections were found in 15 gastropods (2.2% prevalence) representing 5 species; 5 of the infected gastropods were D. laeve. New host records for this nematode are Ventridens collisella and Philomycus carolinianus. Infected gastropods were recovered from June through October 1985 and in May 1986. All 6 sampling locations yielded infected gastropods, suggesting this parasite is widely distributed at the CRC, and is therefore likely to continue to pose a serious threat to exotic ungulates maintained there for conservation and research purposes.

Mycobacterium tuberculosis in zoo and wildlife species.

Tuberculosis caused by Mycobacterium tuberculosis and M. tuberculosis-like organisms has been identified in a wide range of species, including non-human primates, elephants and other exotic ungulates, carnivores, marine mammals and psittacine birds. Disease associated with M. tuberculosis has occurred mostly within captive settings and does not appear to occur naturally in free-living mammals. Mycobacterium tuberculosis probably originated as an infection of humans, but from the zoonotic standpoint, non-human primates, Asian elephants and psittacine birds have the potential to transmit this disease to humans. However, the overall prevalence of disease in these susceptible species is low and documented transmissions of M. tuberculosis between animals and humans are uncommon. Mycobacterium tuberculosis causes progressive pulmonary disease in mammals and a muco-cutaneous disease in parrots. In all cases, the disease can disseminate and be shed into the environment. Diagnosis in living animals is based on intradermal tuberculin testing in non-human primates, culturing trunk secretions in elephants, and biopsy and culture of external lesions in parrots. Ancillary testing with deoxyribonucleic acid probes and nucleic acid amplification, and enzyme-linked immunosorbent assays have been adapted to some of these species with promising results. Additionally, new guidelines for controlling tuberculosis in elephants in the United States of America, and programmes for tuberculosis prevention in animal handlers have been established.

Deaths from exertional myopathy at the National Zoological Park from 1975 to 1985.

A retrospective study was conducted to determine the incidence of deaths from exertional myopathy (EM) in the hoofstock collection at the National Zoological Park (NZP) from 1975 to 1985. The diagnosis of EM was based on history, clinical signs, clinicopathological findings, gross and microscopic lesions. Only bovids, cervids, and equids were included in the study. Data were derived from the medical and pathological records of the NZP. There were 10 cases of EM during this period, but only seven deaths were directly attributable to EM. Five deaths occurred after immobilization. Two deaths occurred after improper transport, but the anesthetic history was not known in these cases. Pathologic changes consistent with EM were incidental findings in an animal killed by dogs, and in two animals euthanized because of suspected infection with Parelaphostrongylus tenuis. Only cervids and bovids were affected. There was no age or sex predilection for developing EM and deaths occurred throughout the year. Deaths associated with immobilization occurred most often after anesthesia with xylazine and etorphine. These are the drugs used most frequently in bovid and cervid anesthesia at the NZP. The overall incidence of deaths from EM after immobilization was 0.25% for the 10-yr period.

Mortality of captive tortoises due to viviparous nematodes of the genus Proatractis (Family Atractidae).

Between September 1982 and January 1984, verminous colitis was diagnosed post mortem in eight red-footed tortoises (Geochelone carbonaria) and three leopard tortoises (Geochelone pardalis) from the reptile collection of the National Zoological Park. This represented 69% of 16 tortoise necropsy accessions for that period. Etiology was determined to be a viviparous pinworm-like nematode of the genus Proatractis (Family Atractidae). Clinical signs were either nonspecific, consisting of anorexia, lethargy, and depression, or were absent. Limited trials with piperazine citrate and fenbendazole appeared to be ineffectual against the parasite and supportive therapy was unsuccessful. Post mortem examination revealed roughening and thickening of the mucosa of the cecum and colon, and in severe cases myriads of tiny (0.5-1.0 cm) nematodes were evident on the mucosal surface. In six tortoises, worms were found also in the small intestine. Histopathologic features in severe cases included mucosal necrosis with parasites and mixed inflammatory cells extending into the tunica muscularis. Focal to diffuse lymphoplasmacytic infiltrates were present consistently in the submucosa of the cecum and colon, and similar but milder lesions occasionally occurred in the small intestine.

Distal extremity necrosis in captive birds.

The necropsy files of the National Zoological Park and Baltimore Zoological Society were reviewed for cases of distal extremity necrosis (DEN) in birds. Nineteen cases of DEN occurred following either trauma or frostbite. Six birds developed an apparently primary type of DEN in which no predisposing factors were obvious clinically. The toes and feet were most commonly involved, and in several cases the beak was also affected. Some pathologic evidence is provided that certain cardiovascular lesions may predispose birds to DEN by compromising circulation of the extremities.

Clinical and pathological findings of a newly recognized disease of elephants caused by endotheliotropic herpesviruses.

The unique clinical and pathological findings in nine Asian (Elephas maximus) and two African (Loxodonta africana) elephants from North American Zoos with a highly fatal disease caused by novel endotheliotropic herpesviruses are described. Identification of the viruses by molecular techniques and some epidemiological aspects of the disease were previously reported. Consensus primer polymerase chain reaction (PCR) combined with sequencing yielded molecular evidence that confirmed the presence of two novel but related herpesviruses associated with the disease, one in Asian elephants and the second in African elephants. Disease onset was acute, with lethargy, edema of the head and thoracic limbs, oral ulceration and cyanosis of the tongue followed by death of most animals in 1 to 7 days. Pertinent laboratory findings in two of three clinically evaluated animals included lymphocytopenia and thrombocytopenia. Two affected young Asian elephants recovered after a 3 to 4 wk course of therapy with the anti-herpesvirus drug famciclovir. Necropsy findings in the fatal cases included pericardial effusion and extensive petechial hemorrhages in the heart and throughout the peritoneal cavity, hepatomegaly, cyanosis of the tongue, intestinal hemorrhage, and ulceration. Histologically, there were extensive microhemorrhages and edema throughout the myocardium and mild, subacute myocarditis. Similar hemorrhagic lesions with inflammation were evident in the tongue, liver, and large intestine. Lesions in these target organs were accompanied by amphophilic to basophilic intranuclear viral inclusion bodies in capillary endothelial cells. Transmission electron microscopy of the endothelial inclusion bodies revealed 80 to 92 nm diameter viral capsids consistent with herpesvirus morphology. The short course of the herpesvirus infections, with sudden deaths in all but the two surviving elephants, was ascribed to acute cardiac failure attributed to herpesvirus-induced capillary injury with extensive myocardial hemorrhage and edema.

Mycobacterial osteomyelitis in captive marsupials.

Mycobacterial osteomyelitis was detected in 3 marsupials exhibited at the National Zoological Park, Washington, DC. One Matschiei's tree kangaroo (Dendrolagus matschiei), 1 Parma wallaby (Macropus parma), and 1 long-nosed rat kangaroo (Potorous tridactylus) were affected. The Parma wallaby had disseminated granulomatosis. Acid-fast organisms were observed in the bone marrow of the wallaby, using the auramine-O-rhodamine fluorescent technique; however, cultures were negative. The tibiotarsal joint of the rat kangaroo contained exudate, with fistulous tracts and necrosis of the articular surface. Granulomas with necrotic centers from this area were positive by auramine-O-rhodamine but were negative on culture. The tree kangaroo had thickening of the right ischium, with a pocket of exudate caudal to the acetabulum. The musculature in the acetabular area was thickened and fibrotic. Mycobacterium avium serotype 15 was isolated from the ischium and liver of this animal.

An epornitic of duck viral enteritis in a zoological park.

An epornitic of duck viral enteritis occurred at the National Zoological Park in the spring of 1975. The disease affected 8 different species of ducks; geese and swan were spared. Diagnosis was based on characteristic pathologic changes, with herpetic inclusion bodies in the epithelium of the digestive tract and liver, and by viral isolation from 4 ducks. An experimental attenuated live-virus vaccine and sanitizing procedures were used to control the epornitic.

Fatal herpesvirus infection in patas monkeys and a black and white colobus monkey.

Fatal herpesvirus infections were diagnosed in 3 patas monkeys and 1 black and white colobus monkey over a 4-week period. Herpesvirus was isolated from 1 patas monkey and from the black and white colobus monkey. Both isolates had growth characteristics similar to Herpesvirus hominis and Herpesvirus simiae. The isolate from the colobus monkey antigenically appeared to be H simiae or H simiae-like, whereas the isolate from the patas monkey could not be conclusively identified with the antisera used. All affected animals were housed in close proximity to rhesus monkeys, the carrier host of H simiae.

Tuberculosis in captive exotic birds.

Avian tuberculosis was studied clinically and pathologically in 137 affected birds from the National Zoological Park during a 7-year period (1969-1975). Twelve of 22 orders exhibited were affected by the disease, and the highest annual mortality was 4% (in 1975). Antemortem diagnosis of early cases of the disease, based on tuberculin testing, and serologic, hematologic, and radiographic studies, was inconsistent and often not conclusive. Pathologically, the diseases primarily involved digestive organs and spleen. There was a spectrum of lesions consisting of nodules of large foamy histiocytes packed with acid-fast bacilli to giant cell-containing granulomas that were often caseous but not cavitated or calcified. Amyloidosis was seen in approximately 20% of the cases. Mycobacterium avium serotype 1 was isolated from 30 tuberculous birds cultured. There was no sex predilection, and most of the affected birds were adults ranging from 1 to 10 years of age.